雌激素和维生素K抗去势成年雌性大鼠骨质疏松的实验研究

Experimental Study of the Estrogen and Vitamin K on Osteoporosis of Ovariectomized Female Rats.

目的寻找一种或一组简便有效,价格低廉的防治骨质疏松(osteoporosis,OP)的方法,以期为防治骨质疏松药物的开发提供实验依据。方法以去卵巢雌性SD大鼠作为绝经早期骨质疏松模型。将50只SD雌性大鼠随机分为5组:卵巢去势(ovariectomized,OVX)组(OVX组,n=10)、卵巢去势+雌激素组(OVX+E组,n=10)、卵巢去势+维生素K组(OVX+vit K组,n=10)和卵巢去势+雌激素+维生素K组(OVX+E+vit K组,n=10)、假手术组(sham组,n=10)。卵巢去势1个月后,分组给药,8周后集中统一处死,取左侧股骨检测骨密度(bone mineral density,BMD),右侧股骨病理学切片进行骨小梁形态计量观察,取右心室血清检测白介素(interleukin,IL)-6、骨钙素(bone gla protem,BGP)和胰岛素样生长因子(insulin-like growth factor,IGF)-1含量。结果①OVX组与sham组比较,骨密度显著下降,血清白介素-6和骨钙素测量值明显升高,而胰岛素生长因子-1显著下降,表明造模成功。②OVX组骨组织病理学切片40倍光镜下骨小梁形态计量观察结果与sham组、OVX+vit K组、OVX+E组和OVX+E+vit K组比较,差异有显著意义。其中,OVX+E+vit K组与sham组最相似。③OVX+E组与OVX组比较,股骨干骺端(松质骨)骨密度显著升高,而股骨中段(皮质骨为主)骨密度升高,差异无显著意义。OVX+vit K组与OVX组比较,股骨中段(皮质骨)骨密度显著升高,股骨干骺端(松质骨为主)骨密度升高,但差异无显著意义。OVX+E+vit K组股骨干骺端和股骨中段骨密度均显著性升高,与sham组最接近。④OVX+E组与OVX组比较,血清白介素-6含量和骨密度低,胰岛素生长因子-1含量升高,差异有显著意义;OVX+vit K组与OVX组比较,血清白介素-6测量值低,骨钙素和胰岛素生长因子-1测量值升高,差异有显著意义;OVX+E+vit K组血清白介素-6,骨钙素和胰岛素生长因子-1含量与sham组比较,最为接近。结论①去卵巢大鼠骨密度及骨形成指标胰岛素生长因子-1下降,骨吸收指标白介素-6、骨吸收和骨形成指标骨钙素升高,表明绝经早期骨代谢处于骨吸收大于骨形成的高转换状态,导致骨质疏松。②雌二醇(estrogen,E_2)水平下降是骨质疏松的一个重要致病因素。病理形态学观察和骨密度及血清白介素-6、骨钙素、胰岛素生长因子-1含量检测表明,维生素K和雌激素防治骨质疏松均有效,二者联合用药的作用明显优于单一用药。

Objective To find a group of simply, effective and cheap method to prevent and remedy osteoporosis（OP） in order to provide the evidence for developing medicines against osteoporosis. Methods Fifty Sprague-Dawley rats were taken as the osteoporosis model of the early menopause. They were randomly divided into 5 groups： ovariectomized group （OVX group, n= 10）, ovariectomize and estrogen treatment group （OVX＋Egroup, n=10）, ovariectomized and vitamin K treatment group （OVX＋vit K group, n= 10）, ovariectomized and estrogen treatment and vitamin K treatment group （OVX＋ E＋vit K group, n=10）, and sham operation group （sham group, n= 10）. These rats were given different medicine according to their groups after a month of ovariectomized operation, then killed together 8 weeks later. The bone mineral density （BMD） of the left femur and morphological observation of bone trabecula for right femur were observed. The content of interleukin-6 （IL-6）, bone gla protein （BGP） and insulin-like growth factor-1 （IGF-1） of blood serum from the right part heart were tested. Results ① Compared with OVX group and sham group, the content of bone mineral density and insulin-like growth factor-1 descend eminently, and the content of interleukin-6 and bone gla protein increased outstandingly. ② The pathological section of bone tissue of OVX group had a significant different compared with sham group, OVX＋vit K group, OVX＋E group, and OVX＋E＋vit K group under the forty times light microscopy. The form of bone tissue of OVX＋E＋vit K group was similar to that of sham group. ③Compared between OVX＋E group and OVX group, the bone mineral density in the femur shaft （loose bone） went up obviously, but the bone mineral density in the middle part of femur（dense bone） went up moderately, which had no statistical difference. The bone mineral density of the middle part of femur（dense bone） increased obviously with no difference with the growth of bone mineral density in femur shaft（ loose bone） between OVX＋vit K group and OVX group. The bone mineral density in the middle part of femur and femur shaft rised dramatically than that of OVX＋E＋ vit K group, which was closed to that of sham group. ④ The content of interleukin-6 and bone gla protein of OVX＋E group was lower than that of OVX group with a significant difference. The content of insulin-like growth factor-1, bone gla protein of OVX＋vit K group went up and interleukin-6 went down compared with OVX group. The content of interleukin-6, bone gla protein and insulin-like growth factor-1 in OVX＋E＋vit K group was closest to that of sham group. Conclusion ①The decrease of contents of bone mineral density and insulin-like growth factor-1 and increase of interleukin-6 and bone gla protein in SD rats indicate that bone metabolism at the early menopause is in a state of high conversion and the reduce of estrogen makes a great contribution to the disease of osteoporosis. ② The decrease of estrogen level plays an important role in the process of osteoporosis. Vitamin K and estrogen have effective precaution against osteoporosis. The combination of estrogen and vitamin K are superior to using estrogen or vitamin K only. factor-1; bone mineral density（BMD）